针对肿瘤相关血管以增加免疫浸润可能增强治疗效果，然而很少有数据可用于评估抗血管生成对肿瘤微环境的影响。我们假设联合靶向免疫检查点的血管生成双重靶向将改善颅内和颅外疾病。我们利用皮下和左心室黑色素瘤模型评估抗PD-1/抗血管内皮生长因子（VEGF）和抗PD-1/lenvatinib（泛VEGF受体抑制剂）组合。进行细胞因子/趋化因子分析和流式细胞术以评估信号和免疫浸润群体。利用体外血脑屏障模型研究颅内治疗对内皮完整性和白细胞越过的影响。抗PD-1与抗VEGF或lenvatinib的组合可改善系统性黑色素瘤小鼠模型的存活率，减少肿瘤生长；治疗增加了TH1细胞因子/趋化因子信号。Lenvatinib减少了肿瘤相关巨噬细胞，但在治疗早期增加了浆细胞样树突状细胞；这种效果在抗VEGF上不明显。Lenvatinib和抗VEGF都导致肿瘤内部的血管减少。虽然抗VEGF促进了体外血脑屏障模型中的内皮稳定，而lenvatinib没有，但两种方案都能促进白细胞越过。联合靶向PD-1和VEGF或其受体可以促进黑色素瘤的抗肿瘤活动，但它们对肿瘤微环境的影响是不同的。这些研究提供了有关双重抗PD-1和抗血管生成组合的见解。

Targeting tumor-associated blood vessels to increase immune infiltration may enhance treatment effectiveness, yet limited data exists regarding anti-angiogenesis effects on the tumor microenvironment. We hypothesized that dual targeting of angiogenesis with immune checkpoints will improve both intracranial and extracranial disease. We used subcutaneous and left ventricle melanoma models to evaluate anti-PD-1/anti-vascular endothelial growth factor (VEGF) and anti-PD-1/lenvatinib (pan-VEGF receptor inhibitor) combinations. Cytokine/chemokine profiling and flow cytometry were performed to assess signaling and immune infiltrating populations. An in vitro blood-brain barrier model was utilized to study intracranial treatment effects on endothelial integrity and leukocyte transmigration. Anti-PD-1 with either anti-VEGF or lenvatinib improved survival and decreased tumor growth in systemic melanoma murine models; treatment increased TH1 cytokine/chemokine signaling. Lenvatinib decreased tumor-associated macrophages but increased plasmacytoid dendritic cells early in treatment; this effect was not evident with anti-VEGF. Both lenvatinib and anti-VEGF resulted in decreased intratumoral blood vessels. Although anti-VEGF promoted endothelial stabilization in an in vitro blood-brain barrier model while lenvatinib did not, both regimens enabled leukocyte transmigration. Combined targeting of PD-1 and VEGF or its receptors promotes enhanced melanoma anti-tumor activity, yet their effects on the tumor microenvironment are quite different. These studies provide insights into dual anti-PD-1 and anti-angiogenesis combinations.